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Naturwissenschaften (2004) 91:5165DOI 10.1007/s00114-003-0493-5
R E V I E W
Jens Amendt Roman Krettek Richard Zehner
Forensic entomology
Published online: 16 January 2004 Springer-Verlag 2004
Abstract Necrophagous insects are important in thedecomposition of cadavers. The close association betweeninsects and corpses and the use of insects in medicocrim-
inal investigations is the subject of forensic entomology.The present paper reviews the historical background ofthis discipline, important postmortem processes, anddiscusses the scientific basis underlying attempts todetermine the time interval since death. Using medicaltechniques, such as the measurement of body temperatureor analysing livor and rigor mortis, time since death canonly be accurately measured for the first two or three daysafter death. In contrast, by calculating the age ofimmature insect stages feeding on a corpse and analysingthe necrophagous species present, postmortem intervalsfrom the first day to several weeks can be estimated.These entomological methods may be hampered by
difficulties associated with species identification, butmodern DNA techniques are contributing to the rapid andauthoritative identification of necrophagous insects. Otheruses of entomological data include the toxicologicalexamination of necrophagous larvae from a corpse toidentify and estimate drugs and toxicants ingested by theperson when alive and the proof of possible postmortemmanipulations. Forensic entomology may even help ininvestigations dealing with people who are alive but inneed of care, by revealing information about cases ofneglect.
Introduction
With about one million described species, insects comprise
the largest metazoan class (Price 1997). They are found inalmost all habitats. One such habitat, providing anexcellent food source for a more or less specialized insectcommunity, is a vertebrate corpse (Anderson and Cervenka2002). About 400 insect species have been found on a pigcadaver during its various stages of decay (Payne 1965).
In addition to their ecological importance in decompo-sition, such insects may represent important tools incriminal investigations (Erzinclioglu 1983; Catts and Goff1992), allowing the time at which a dead body wascolonized to be estimated (Greenberg 1991). In particular,blowflies (Calliphoridae), among the first colonizers ofcadavers, may serve as a biological clock in measuring the
time of death for two or more weeks. Such an entomo-logically-based estimate may be far more precise than themedical examiners, which is limited to about a day or twopostmortem (Greenberg and Kunich 2002). Thereforeforensic entomology, defined as the use of insects andother arthropods, such as mites, in medicocriminal inves-tigations (Hall 2001), is becoming an important field inlegal medicine. The present article describes the historicaldevelopment of forensic entomology, the methods, currentfields of research and future trends.
Retrospective
Insects were first used in a forensic context in thirteenth-century China (McKnight 1981). A farmer had beenkilled in a rice field with a sharp weapon. All the suspectswere called together and were told to place their sickleson the ground. No obvious evidence could be seen, butone sickle attracted numerous blowflies, apparentlybecause of invisible traces of blood on the blade. Theowner of the sickle, when confronted with this entomo-logical evidence, confessed to the killing.
During medieval times, the correlation between mag-gots on a cadaver and the oviposition of adult flies was
J. Amendt ()) R. ZehnerZentrum der Rechtsmedizin, J.W. Goethe-Universitt,Kennedyallee 104, 60596 Frankfurt, Germanye-mail: amendt@em.uni-frankfurt.deTel.: +49-69-63017571Fax: +49-69-63015882
R. KrettekFB 19, FG kologie, Universitt Kassel,Heinrich-Plett-Strasse 40, 34132 Kassel, Germany
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promoted by the internal chemical breakdown of cells andreleased enzymes as well as by the activity of bacteria andfungi, from both the intestine and the external environ-ment (Knight 1991; Clark et al. 1997; Introna andCampobasso 2000). The body temperature decreases(algor mortis) and the skin colour reddens (livor mortisor lividity), which is generally evident at about 2 hpostmortem. This is due to the gravitational pooling of
blood in dependent body parts. After a few hours thecolour changes from red to purple as oxygen graduallydissociates from the haemoglobin of the red blood cells.By 46 h after death, lividity is fixed because the fat inthe dermis solidifies in the capillaries. Another sign ofdeath is the stiffening of the muscle fibres due to thebreakdown of glycogen and the accumulation of lacticacid (rigor mortis). This is first noticeable in the facialmuscles 23 h postmortem and reaches its maximum after24 h. The duration of rigor mortis depends on themetabolic state at death and on various factors such asbody size and surrounding temperature. Later skinslippage, the loosening of the epidermis from the
underlying dermis, occurs and hair and nails are easilyremoved. Large quantities of putrefaction gases cause thephysical distortion of the body. Hydrogen sulphide (H2S)reacts with haemoglobin and forms a green pigmentwhich initially shows up the superficial blood vessels, butlater may also be seen as a green coloration in thegastrointestinal region and those portions of the bodywhere livor mortis was most marked. All these signsoccur within the first 7296 h after death (Henge et al.1995, 2000a, 2000b). However, the precise rate ofpostmortem decay is affected by a wide range of variablesassociated with the corpse itself and the surroundingenvironment. Moreover, after the temperature of the body
has equilibrated with that of the environment andfollowing the initial putrefaction, no reliable estimationof the postmortem interval (time since death) is possible.Subsequently, therefore, insects found on the bodyprovide an important source of information.
Insects and death
Insects are attracted to a body immediately after death,often within minutes (Erzinclioglu 1983; Smith 1986;Anderson and VanLaerhoven 1996; Haskell et al. 1997;Anderson 2001). However, oviposition may not occur.Many taxa which appear very early at a death scene arelate colonizers or even non-necrophagous species.
According to Smith (1986), four ecological categoriescan be identified in a carrion community:
1. Necrophagous species, feeding on the carrion.2. Predators and parasites of necrophagous species,
feeding on other insects or arthropods. This groupalso contains schizophagous species, which feed on thecarrion at first, but may become predaceous in laterlarval stages.
3. Omnivorous species such as wasps, ants and somebeetles feeding both on the corpse and its colonizers.
4. Other species, such as springtails and spiders, whichuse the corpse as an extension of their environment.
For the purposes of forensic entomology, the first twogroups are the most important. They include mainlyspecies from the orders Diptera (flies) and Coleoptera(beetles) (see Table 2). The succession on corpses can bedivided into different waves over the various stages ofdecay, although this has been debated (Schoenly and Reid1987). Nevertheless, since the attractiveness of a decayingbody differs between necrophilous insects, changes overtime and the colonization of the corpse will occur in apredictable sequence (see Fig. 1).
Blowflies, are typically the first colonizers, attracted tothe carrion by the odour produced during decomposition(Wall and Warnes 1994; Fisher et al. 1998; Anderson2001), even over large distances (Braack 1981; Erzincli-oglu 1996). Besides olfactory stimuli, vision, colour andthe presence of other conspecific insects on the dead bodyall play a role (Hall 1995; Hall et al 1995; Wall and Fisher2001). The presence of ammonia-rich compounds andhydrogen sulphide are important stimulants for oviposi-tion, as well as moisture, some pheromones, and tactilestimulants (Ashworth and Wall 1994; Fisher et al. 1998;Anderson 2001). Female Diptera do not oviposit indehydrated or mummified tissue, as eggs and larvae needmoisture for successful development (Introna and Cam-
Table 2 Selection of insects of forensic importance
Order/Family Important genera
COLEOPTERA/BEETLES
Cleridae (Checkered beetles) NecrobiaDermestidae (Larder beetles) Attagenus, DermestesGeotrupidae (Dung beetles) GeotrupesHisteridae (Clown beetles) Hister, SaprinusSilphidae (Carrion beetles) Necrodes, Nicrophorus, Silpha
Staphylinidae (Rove beetles) Aleochara, CreophilusDIPTERA/FLIES
Calliphoridae (Blowflies) Calliphora, Chrysomya,Cochliomyia, Lucilia, Phormia
Drosophilidae (Fruit flies) DrosophilaEphydridae (Shore flies) DiscomyzaFanniidae (Latrine flies) FanniaHeleomyzidae (Sun flies) Heleomyza, NeoleriaMuscidae (House flies) Hydrotaea, Musca, Muscina,
OphyraPhoridae (Scuttle flies) Conicera, MegaseliaPiophilidae (Skipper flies) Piophila, StearibiaSarcophagidae (Flesh flies) Liopygia, SarcophagaSepsidae (Black scavenger flies) Nemopoda, ThemiraSphaeroceridae (Small dung flies) LeptoceraStratiomyidae (Soldier flies) Hermetia, SargusTrichoceridae (Winter gnats) TrichoceraLEPIDOPTERA/BUTTERFLIES
Tineidae (Clothes moths) Tineola
HYMENOPTERA/WASPS
Ichneumonidae (Ichneumon wasps) AlysiaPteromalidae (Fly wasps) Nasonia, Muscidifurax
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pobasso 2000). Oviposition first occurs at the orifices orwounds of the corpse. The size of the carcass seems toaffect its attractiveness, at least to species of the flyfamilies Calliphoridae and Sarcophagidae (Nuorteva1959b; Davies 1990; Erzinclioglu 1996; Povoln andVerves 1997). However, not all necrophagous insectsprefer larger carcasses; some species oviposit preferen-tially on smaller animals such as rodents or even snails.Several factors restrict the colonization of a corpse, suchas its burial (Mann et al. 1990). Most Diptera are not able
to colonize bodies buried deeper than 30 cm (Introna andCampobasso 2000; Campobasso et al. 2001); however,exceptionally, groups such as the Phoridae may be foundin buried coffins (Schmitz 1928; Stafford 1971; Smith1986; Anderson 2001). Burial, therefore, will influencethe time required for insects to reach the carcass as well asthe species composition of the necrophagous fauna(Payne et al. 1968; Rodriguez and Bass 1985; VanLaer-hoven and Anderson 1999; Campobasso et al. 2001;Bourel et al. 2004). Such a delay may not only occur inburied corpses, but also in those that are covered orwrapped (Goff 1991) or in cadavers found at indoorscenarios (authors unpublished data).
Studies on animal carcasses (Fig. 2) have demonstratedthat species composition and insect succession on acadaver vary with respect to the geographical region andthe season (Bornemissza 1957; Reed 1958; Payne 1965;Goddard and Lago 1985; Introna et al. 1991; Andersonand VanLaerhoven 1996; Richards and Goff 1997;Anderson 2001; Arnaldos et al. 2001; Carvalho andLinhares 2001; Grassberger and Frank 2003a; Watson andCarlton 2003). Data collected for a particular region orarea should be used with caution when determining timeof death in another region. Even local characteristics of
the death scene, like the ecology of the area or the degreeof sun exposure, can alter the pattern of insect coloniza-tion (Smith 1986; Shean et al. 1993; Erzinclioglu 1996).Some insect species are found in both urban and ruralareas, while others are very specific to a certain habitat(Catts and Haskell 1990). Since species commonlyconsidered as rural species have also been collected inurban regions, care must be used in determining whetherremains have been moved based on entomologicalevidence alone (Anderson 2001; Grassberger and Frank
2003a). For example, the blowfly Calliphora vomitoria,usually considered to be a rural species, has also beenfound in residential apartments (C. Reiter, personalcommunication; authors unpublished data).
Estimating time since death
When human remains are found days, weeks, or evenlonger after death, body temperature, and conditions suchas rigor mortis or livor mortis are no longer appropriatefor estimating time since death. In such cases, insects mayprovide important indications of the postmortem interval
(PMI). The ages of insect immature stages found on adead body can provide evidence for the estimation of aminimum PMI ranging from 1 day up to more than1 month, depending on the insect species involved and theclimatic conditions at the death scene. However, thisperiod will not always match the exact PMI. Moreover,the infestation of live vertebrates by flies, called myiasis(Zumpt 1965), is not only a veterinary problem but hasbeen reported in humans (see, e.g., Greenberg 1984;Erzinclioglu 1996; Sherman 2000), and should be kept in
Fig. 1 Succession of adult arthropods on human cadavers in east Tennessee (during spring and summer); adapted from Rodriguez and
Bass (1983) and Hall (2001)
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mind when using entomological evidence in a death sceneinvestigation.
Exact species identification of insect samples is thefirst essential step in estimating the age of the larvae
found. Insect larvae differ in growth rates and biology.Larvae of Lucilia sericata, for instance, grow faster at25C than larvae of Calliphora vicina; blowflies usuallydeposit eggs on a corpse (but see below) while fleshfliesare larviparous. These examples demonstrate that thesame stage of development of larvae found on a corpse donot necessarily indicate the same age or the same time ofcolonization. For estimating the minimum PMI, the age ofthe immature larval stages must be determined. Variousprocedures for estimating their age exist, but all are basedon the fact that the rate of development depends on the
ambient temperature (see Table 3, Fig. 3). Measuring thelength or the dry weight of the oldest larva may reveal itsage by comparing it with reference data (e.g. Reiter 1984;Nishida et al. 1986; Davies and Ratcliffe 1994; Wells and
LaMotte 1995; Grassberger and Reiter 2001, 2002a,2002b). Another approach, known as thermal summation(Wigglesworth 1972), is the accumulation of degree hours(ADH) or degree days (ADD). According to Greenbergand Kunich (2002) it is assumed that the relation betweenthe rate of development and temperature is linear in themid-range of a sigmoidal curve, with an upper and alower threshold below which development ceases. Thetotal amount of heat required, between the lower andupper thresholds, for an insect to develop from the time ofoviposition to the time of hatching is calculated in units
Fig. 2AD Decomposition of a pig carcass in a forest in Germanyduring a period of 42 days (mid-June until end of July); meantemperature 19.5C (minimum 10.3C, maximum 32.9C); A
postmortem interval of 2 days, B PMI of 14 days, C PMI of32 days, D PMI of 42 days
Table 3 Development data forCalliphora vicina(from Ander-son 2000)
15.8 0.004CTime to reach stage (h)
20.6 0.03CTime to reach stage (h)
23.3 0.02CTime to reach stage (h)
Stage Min Max Min Max Min Max1st instar 41.41.2 46.70.6 22.50.2 36.04.0 210 29.55.22nd instar 83.010.0 88.312.7 57.06.0 57.06.0 45.00 52.003rd instar
(feeding stage)128.09.0 146.015.6 84.010.0 93.50.5 77.00 85.08.0
Prepupal 228.03.3 257.09.6 155.54.2 162.51.1 146.00 173.00Pupal stage 294.04.7 440.342.1 213.04.5 233.00.9 202.85.8 279.022.5Adult 719.76.0 874.620.7 514.83.7 572.010.0 454.06.0 499.57.5
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called degree days or degree hours (Baskerville and Emin1969; Allen 1976). Laboratory data are obtained bysumming the number of hours from egg to adult and thenmultiplying by the temperature, after subtracting thetemperature of the lower developmental threshold. Hence,degree-days or -hours are the accumulated product of timeand temperature between the developmental thresholdsfor each day. Each developmental stage has its own totaldevelopmental requirement and each species requires adefined number of degree-days to complete its develop-ment. This fact helps us to predict the time when a certain
developmental stage will be reached. Numerous papersdescribe ADH/ADD values and thresholds for forensical-ly important insects, mainly blowflies and fleshflies (e.g.Kamal 1958; Greenberg 1991; Byrd and Butler 1998;Anderson 2000; Marchenko 2001; but see Ames andTurner 2003; Kaneshrajah and Turner 2004). The useful-ness of these methods depends on the thermal history ofthe immature stages and, to be used, they thereforerequire the evaluation of temperature data at the deathscene before the body was found. Hence, multipletemperature measurements have to be taken at the crimescene, at the body as well in the maggot masses on thecadaver. These data should be compared with data
obtained from the nearest weather station. If these datasets are reasonably similar, the weather station data cansimply be used to extrapolate temperatures at the crimescene (Greenberg and Kunich 2002). Alternatively, ifmore substantive differences exist, the weather stationtemperature data have to be corrected for the averagedifference using mathematical methods such as linearregression, prior to extrapolation of the expected temper-atures at the crime scene. However, when using estimatesof ambient temperature to calculate development rates, itshould be noted that these data are not necessarily
representative of the temperatures experienced by theinsects in the corpse. Large numbers of maggots maycreate a so-called maggot mass effect which, accordingto the metabolic and feeding rate of these immatureinsects, can generate a temperature substantially higherthan ambient (Wells and LaMotte 1995). Moreover,development data obtained in different geographicalregions may not be comparable (Greenberg 1991; Grass-
berger and Reiter 2001). Populations of Lucilia sericatain Russia, for example, may have different minimumthreshold temperatures than populations in the UK, andmay therefore develop at different rates under the sametemperature conditions.
Knowing the chronology of insects colonizing carrionin a certain area (see also the section titled Insects anddeath), analysis of the fauna on a carcass can be used toestimate the time elapsed since death (Goff and Flynn1991; Anderson 2001). A simple succession model can beused when estimating both the age of a larva and the timeinterval between death and the insects arrival on the body(Wells et al. 2001b). Succession data have been used to
calculate a PMI up to 52 days (Schoenly et al. 1996) and,if there are adequate data, may be applied to a muchlonger time interval (authors unpublished data).
DNA analysis in forensic entomology
In forensic entomology, information is essential not onlyon the development stages of the insects found on thebody, but also on their identity. Morphological methodsare usually used (Schumann 1971; Smith 1986; Povolnand Verves 1997). However, these techniques requirespecialized taxonomic knowledge. Although identifica-
tion keys are available, only a few experts are able toidentify the larvae of forensically relevant insects tospecies level. Furthermore, for some groups of insects(e.g. Sarcophagidae) differentiation at the larval stagesusing morphological criteria is still not possible. Time-consuming rearing of the larvae to adults for identifica-tion may delay the criminal investigation or causesignificant problems when rearing fails. Under thesecircumstances, species identification based on geneticexamination is an option. PCR amplification of suitableregions of the genome, sequence analysis of the ampli-cons obtained, and alignment of the data with referencesequences is the usual and recommended method.
For sequence analysis, DNA has to be isolated fromthe specimen. This can be done by various establishedmethods including phenol/chloroform extraction (Sam-brook and Russel 2001), CTAB extraction (Stevens andWall 1996), Chelex extraction (Junqueira et al. 2002), orusing commercial extraction kits, such as the QiAmpTis-sue Kit (Qiagen, Hilden, Germany; Wells et al. 2001a) orDNAzol (Molecular Research Center, Cincinnati, Ohio,USA; Wallman and Donellan 2001). DNA extractionfrom specimens collected at a crime scene is usuallysuccessful. However, with museum specimens, whichmay represent reference species, an extraction of typable
Fig. 3 Growth curves for the blowfly Protophormia terraenovae,showing the time required to reach the larval, pupal and adult stagesat 15, 20, 25, 30 and 35C; areas between lines represent identicalmorphological stages, e.g. pupa (Grassberger and Reiter 2002a)
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DNA is not always successful due to damage of the DNAduring storage (Pbo 1989; Pbo et al. 1989) or to theinfluence of the ethyl esters used for killing the insects(Dillon et al. 1996).
After DNA extraction, PCR and subsequent sequenceanalysis can be performed, either as described by theauthors cited above or by evaluating new primers forspecific gene regions of interest. Frequently investigated
genes are the subunits I and II of the cytochrome oxidase,ND5, ND1, 12S and 16S DNA (mitochondrial encoded)as well as 28S, ITSI and II DNA (nuclear encoded). Foran overview, see Simon et al. (1994), Loxdale and Lushai(1998) and Caterino et al. (2000). To date, mitochondrialgenes in particular have been analysed; sequence infor-mation of the complete mitochondrial genome is availablefor about 400 species (deposited at GOBASE, http://megasun.bch.umontreal.ca/gobase/; Shimko et al. 2001).
Flies are the most important insects in forensicentomology and therefore genetic research has focusedon Diptera (Sperling et al. 1994; Wells and Sperling 1999;Malgorn and Coquoz 1999; Wells and Sperling 2000,
2001; Wells et al. 2001a, 2001b; Stevens and Wall 2001;Wallman and Donellan 2001, Harvey et al. 2003). In mostof these investigations the use of the genes of subunit Iand/or II of the mitochondrial encoded gene for cyto-chrome oxidase, a part of the respiratory chain within themitochondrial membrane, has been examined.
When a sequence of an unknown insect matches areference sequence, it can be concluded that these twotaxa are identical or at least belong to the same speciescomplex. If not, different species can be assumedbecause, in most cases, considerable differences betweenspecies can be observed (Table 4). However, wheredifferences occur, information about the intraspecific vs
the interspecific variation is necessary in order to evaluatethese differences. Wells and Sperling (1999, 2001)demonstrated, by examining COI and COII sequences,that the blowflies Chrysomya rufifacies and Chrysomyaalbicepsexhibit less than 1% intraspecific and about 3%interspecific differences. However, the authors also statethat these data have to be seen as preliminary because anoverlap of intraspecific and interspecific sequence vari-
ation cannot be excluded based on the data currentlyavailable. Similar observations have been reported for thefleshflies Sarcophaga argyrostoma and S. crassipalpis[intraspecific variation 1%, interspecific variation about3%, respectively (Wells et al. 2001a)] and the blowfliesCalliphora vicina and C. vomitoria [intraspecific vari-ability of less than 1%, interspecific variability of about5% in the COI (Vincent et al. 2000) and COII subunits
(authors unpublished data)].Since intraspecific variation will mostly be smallerthan interspecific variation, unambiguous identification atthe species level may be possible. However, carefulinterpretation should be employed until more data areavailable. The need for caution is also demonstrated bythe following observations: Stevens and Wall (1996)examined the mitochondrial encoded 12S rRNA, COI andCOII sequences of the two blowflies Lucilia cuprina andL. sericata originating from different geographical re-gions throughout the world. Lucilia cuprina specimensfrom Hawaii, which exhibit a clear affiliation to thisspecies on the basis of morphological characters, were
assigned to L. sericata rather than to L. cuprina on thebasis of the mitochondrial sequence data. This may reflecthybridization between L. sericata and L. cuprina inHawaii and this is supported by analysis of the nuclearencoded 28S rDNA sequence, which was identical to thatofL. cuprinafrom various other locations outside Hawaii(Stevens et al. 2002)
In addition, unexpected variability between two spec-imens ofC. vomitoriafrom different geographical origins(the USA and the UK) was found within the D1D7region of the nuclear encoded 28 s rRNA sequences(Stevens and Wall 2001), although the 28S gene is knownto be quite conservative. These differences between two
individuals exhibited the same values as between those ofC. vicina and C. vomitoria(both from the UK).Nevertheless, species analysis based on DNA sequence
data appears to be promising. Examining the intraspecificvariability of individuals of the same species collectedfrom distant locations showed a relatively low variabilitycompared with interspecific differences (Stevens andWall 1996, 2001). However, studies on the intraspecific
Table 4 Pairwise percent sequence differences for a 386 bp region of COI of selected Sarcophagids compared to Drosophila yakuba(fromZehner et al. 2004b)
1 2 3 4 5 6 7 8 9 10 11 12
1 Sarcophaga carnaria 2 Sarcophaga subvicina 4.7 3 Sarcophaga variegata 2.7 4.1 4 Parasarcophaga albiceps 8.4 8.1 8.1 5 Bercaea africa 9.5 9.1 8.4 9.8 6 Liopygia argyrostoma 6.8 6.1 7.1 7.8 7.4 7 Liopygia crassipalpis 8.4 8.1 7.8 8.1 6.1 6.1 8 Liosarcophaga teretirostris 8.4 7.8 8.8 7.8 8.8 8.1 7.8 9 Liosarcophaga tibialis 9.1 8.1 8.8 6.4 9.1 7.1 8.8 7.1
10 Pandelleana protuberans 9.8 8.8 8.8 7.8 10.1 9.5 9.5 8.8 9.1 11 Thyrsocnema incisilobata 8.4 7.1 7.8 8.8 9.5 7.4 8.8 9.5 8.4 10.5 12 Helicophagella melanura 9.5 8.4 8.4 8.1 8.8 6.8 7.4 8.4 8.1 8.4 7.1 13 Drosophila yakuba 13.5 14.2 13.5 14.5 12.2 11.8 12.5 13.9 15.2 15.2 14.5 14.2
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variability of a much larger number of individuals arenecessary in order to provide a more solid basis forspecies identification.
In general, with sequence analysis, such as for COI,the maximum molecular data are obtained. By applyingother techniques such as PCR-RFLP, where specificrestriction patterns are produced due to different restric-tion sites in the sequences (Sperling et al. 1994; Malgorn
and Coquoz 1999; Vincent et al. 2001; Schrder et al.2003), only limited information about a small regionwithin the sequence, such as the restriction site within theamplicon, is achieved. In cases of intraspecific variationat a certain restriction site, this may lead to falseexclusions because an unknown restriction pattern mayoccur, although the other parts of the sequence areidentical. In addition, the technique of random amplifiedpolymorphic DNA fingerprinting (RAPD, Williams et al.1990; Benecke 1998) or single strand conformationpolymorphism (SSCP, Gasser and Chilton 2001; Rathand Ansorg 2000) can only be used in direct site-to-sitecomparisons, because it is not possible to generate data
which are comparable from one laboratory to another.Although these techniques offers fast results they cannotbe used for direct and unambiguous species determina-tion.
The technique of sequence analysis, in particular, hasbecome very popular in recent years. Analysis kits,computer-based sequencers and software-aided analysisof the sequences generated make sequence analysisrelatively easy. Moreover, many companies now offersequence analysis for a relatively small charge. Sequenceanalysis may be the method of choice for speciesdetermination.
The analysis of human DNA extracted from maggots is
another important application of molecular tools (Wells etal. 2001b; Clery 2001). This kind of analysis may becomeimportant in cases where the source of the maggots foodis disputed, when only maggots but no corpse is found atthe scene of possible murder, or where an alternative foodsource is present at the scene. By detecting human DNAin the digestive tract of the maggots, it may be demon-strated that they have fed on a human cadaver. Byanalysis of individual-specific DNA (mitochondrial d-loop, STR) a maggot can be assigned to a specific corpse(Zehner et al. 2004a).
EntomotoxicologyLarvae which feed on corpses may sequester drugs andtoxicants which had been ingested by the deceasedperson. Analysis of carrion-feeding insects, to detecttoxic substances and to investigate the effects on insectdevelopment, is known as entomotoxicology (Goff andLord 2001). Bodies in a state of advanced decompositionor that are skeletonized may be difficult to examine fortoxicologically relevant substances due to the lack ofappropriate sources such as tissue, blood or urine. Instead,analysis of the insects encountered may enable toxico-
logical assessment of the cause of death (Nolte et al.1992; Goff and Lord 1994, 2001; Introna et al. 2001;Campobasso et al. 2004; but see Tracqui et al. 2004).After maceration of the larvae, analyses such as thin-layerchromatography (TLC), radioimmunoassay (RIA), gaschromatography (GC), gas chromatography/mass spec-trometry (GC/MS), or high-performance liquid chroma-tography/mass spectrometry (HPLC/MS) may be
performed (Gagliano-Candela and Aventaggiato 2001;Goff and Lord 2001). This can also be applied to adultinsects or even remnants of larval and puparial shells,which are often found at the death scene, even afterseveral years (Miller et al. 1994; Bourel et al. 2001a,2001b).
The detection of mercury in the larvae of variousspecies of blowfly reared on tissues containing knownconcentrations of this metal was described by Nuortevaand Nuorteva (1982). Their study was based on a case ofan unidentified female corpse found in an advanced stageof decomposition in a rural area of Finland (Nuorteva1977). Fly larvae were collected from the corpse, allowed
to complete their development and the emerged adultsanalysed for mercury. The low concentration of mercurydetected in the flies indicated that the victim was from anarea relatively free of mercury pollution and not from thearea where her body was found. These findings drew theattention of the police to a certain area, enabling thesuccessful identification of the victim and resolution ofthe case. Kintz et al. (1990) demonstrated that toxicolog-ical data from Diptera larvae seem to be more reliablethan those from cadaver tissues. Benzodiazepines, barbi-turates and tricyclic antidepressants were detected incalliphorid larvae collected from a corpse 67 dayspostmortem. Goff et al. (1997) analysed antidepressant
drugs from maggots and empty puparia of Diptera; Milleret al. (1994) showed the presence of these drugs in emptyDiptera puparia (Phoridae), cast beetle exuviae (Der-mestidae) and even in faecal material of beetles (Der-mestidae). The usefulness of entomotoxicologicalmethods has been demonstrated in experimental (e.g.Introna et al. 1990; Sadler et al. 1997a; Goff et al. 1997;Hedouin et al. 2001; Pien et al 2004) as well asmiscellaneous case studies (e.g. Beyer et al. 1980; Kintzet al. 1994; Sadler et al. 1995).
Ingested drugs or toxicants may influence the devel-opment of the necrophagous insects (OBrien and Turner2004). Goff et al. studied the effects of cocaine (Goff etal. 1989) and heroin (Goff et al. 1991) on the rate ofdevelopment in Sarcophagidae and demonstrated thatmaggots ofBoetterisca peregrinadevelop more rapidly ifreared on the liver or spleen of rabbits which had beenkilled by a lethal dose of cocaine or heroin. Thisillustrates the potential impact of drugs when estimatingpostmortem intervals by calculating the rate of develop-ment. Bourel et al. (1999) showed that an underestimationof the postmortem interval up to 24 h is possible if thepresence of morphine in tissue is not considered whencalculating the development time ofLucilia sericata. In asuicide case where the pesticide malathion had been used,
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the development stages of two blowflies, Chrysomyamegacephalaand C. rufifacies, found on the dead body,indicated a minimum postmortem interval of 5 days,although the victim had last been seen alive 8 days priorto the discovery of his body (Gunatilake and Goff 1989).The authors concluded that malathion in the tissuesdelayed the colonization by insects for several days.
The absence of a drug in larvae may not indicate that
the drug was not present in the food source (Sadler et al.1997a, 1997b, 1997c). The toxicological analysis ofCalliphora vicinalarvae reared on a substrate containingfour benzodiazepines yielded a negative result for therapidly eliminated loprazolam, although bromazepam anddiazepam were detected (Sadler et al 1997b). Larval drugand food source concentrations differed in an unpre-dictable ways and were found not to be useful forquantitative calculations.
These examples demonstrate that insects found oncorpses can be used in toxicological analyses, but alsoillustrate the risk of calculating an incorrect postmorteminterval because of a modified rate of development of the
immature stages. Further research should focus on thebioaccumulation and metabolism of drugs in necropha-gous insects and their effects on the rate of development.
Future trends in forensic entomology
The precise estimation of PMI is the most important goalof forensic entomology by refining the techniques used.Developmental and succession data, consideration of agreater number of geographical regions and a range ofdeath scene scenarios are essential. Moreover there areseveral parameters which need further attention.
It is important to consider factors that might alter thetime of oviposition, such as covering corpses withbranches or tight wrapping with blankets, carpets orplastic bags, and indoor placement, because these factorsmay delay initial oviposition (Higley and Haskell 2001).Seasonal influences, such as cold and rainy weather, mayinhibit or even prevent fly activity and delay oviposition(Erzinclioglu 1996). However, Faucherre et al. (1999)observed flying as well as ovipositing Calliphora vicinaunder extreme conditions in the Swiss Alps, colonizing acorpse in a 10-m deep cave at a temperature of about 5C.The generally accepted assumption that activity ofnecrophagous flies ceases below an air temperature of10C (Williams 1984) or even 12C (Smith 1986;Erzinclioglu 1996) may be questionable (see also Deonier1940; Nuorteva 1965). However, the case described byFaucherre et al. (1999) occurred at an altitude of 1,260 mand therefore a cold-adapted population ofC. vicinamayhave been involved.
Blowflies usually show peaks of oviposition activity inthe early afternoon (Nuorteva 1959a; Baumgartner andGreeenberg 1984, 1985; Greenberg 1990). These insectsare not active at night and generally do not lay eggsduring nighttime (Greenberg 1985). A postmortem inter-val estimation based on that assumption has to consider
the possibility that a corpse which was found about noonand was infested by recently hatched maggots, could havebeen deposited there in the late evening of the previousday. Hence, fly eggs detected on a corpse during the nightwould lead to the conclusion that death occurred duringthe previous day or earlier (Nuorteva 1977). Greenberg(1990) presented the first experimental evidence ofnocturnal oviposition by three forensically important
blow flies,Calliphora vicina,Phormia reginaandLucilia(Phaenicia) sericata. On the other hand, Tessmer et al.(1995) reported that blowflies fail to lay eggs at nightboth in urban (with lighting) and rural dark habitats.However, Singh and Bharti (2001) supported the findingsof Greenberg (1990). Hence nocturnal oviposition is apossibility and should be taken into consideration.
Diapause, the period during which growth and devel-opment of insects is suspended, is still a challenge for theforensic entomologist (see also Ames and Turner 2003).Depending on the insect taxa, the major influences onlarvae or pupae are photoperiod and temperature. Declin-ing day length and/or decreasing temperatures indicate
approaching winter and induce diapause, preventingdevelopment under unfavourable environmental condi-tions. In many forensically important blowflies, diapauseis under maternal control and exposure of females to shortday lengths induces diapause in the offspring (Vino-gradova 1991). Species with a large geographical rangehave to face changes in day length throughout the year.The critical day length which induces diapause will belonger in populations from a northern range than insouthern populations (McWatters and Saunders 1998).The forensic entomologist working in a temperate regioninvestigating a sample of dead maggots collected from acorpse during late September has to consider the possi-
bility that these maggots had already entered diapause.Besides day length, temperature may also influence theincidence of diapause (Vinogradova and Zinovjeva 1972).Unlike photoperiod, temperature is not a noise-free signal,as it is subject to considerable variation both within andbetween years (McWatters and Saunders 1998). Increas-ing constant temperature is known to reduce the incidenceof diapause in forensically important Dipteran species,such as Liopygia argyrostoma (Saunders 1975), Pro-tophormia terraenovae (Vinogradova 1986) and Calli-phora vicina(McWatters and Saunders 1998).
The duration of diapause is another important param-eter. McWatters and Saunders (1998) showed that in C.vicina kept at temperatures of 15C and 20C, respec-tively, diapause was terminated in most larvae within30 days. However, the diapause ended earlier in larvaewhose parents had been kept at 20C than those whoseparents had been kept at 15C. These observations shouldbe a caveat for the forensic entomologist and points to theneed for further studies on other species.
Competition may affect development and growth ofthe larvae. Smith and Wall (1997a, 1997b) presented datawhich indicate that the larvae of Lucilia sericata incarcasses experience significant levels of competition andthat the intensity of this competition may be sufficient to
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reduce the numbers of adult L. sericata able to emergesuccessfully.
Reiter (1984), Smith (1986) and Erzinclioglu (1990)pointed to another factor which could complicate theestimation of the postmortem intervalprecocious eggdevelopment in flies. In some female flies, eggs may beretained in the oviduct, having been fertilized as they passthe spermathecal ducts in advance of the act of oviposi-
tion (Wells and King 2001). In cases where a suitableoviposition site is not available, the eggs may remaininside the fly until they have completed embryonicdevelopment. It has been reported for several species ofthe tribe Calliphorini, including the forensically importantCalliphora vicina, that the larva hatches from such eggsimmediately following oviposition (Erzinclioglu 1990;Wells and King 2001). Precocious eggs are more likely tobe found in bluebottles (Calliphora spp.) than in otherlineages of carrion-feeding blowflies and the proportionof wild flies carrying an egg that is about to hatch can bequite high (Wells and King 2001).
Parasitoid larvae feed exclusively on other arthropods,
mainly insects, resulting in the death of the parasitoidshost (Godfray 1994). The majority of parasitoids areeither members of the order Hymenoptera or Diptera,representing an extremely diverse group and constitutingabout 8.5% of all described insects (LaSalle and Gauld1991; Godfray 1994). They also attack necrophagous taxaand therefore could appear on carrion. Fabritius andKlunker (1991) listed 83 parasitoid species, mainlywasps, which attack the larval and pupal stages ofsynanthropic Diptera in Europe. There are few reports onthe use of parasitoids in forensic entomology (Smith1986; Haskell et al. 1997; Amendt et al. 2000; Andersonand Cervenka 2002; Grassberger and Frank 2003b). The
life-cycles of the common parasitoid species are known(e.g. Geden 1997) and, even if the adults have alreadyemerged and left the host, the pupal exuviae of theparasitic wasps can be identified for a long timeafterwards (Geden et al. 1998; Carlson et al. 1999). Theparasitoid developmental times can then be calculated andadded to the time of development of the blowfly host.Pupal parasitoids of blowflies may play an especiallyimportant role in the estimation of the postmortem periodbecause their time of attack is often restricted to a small,well-defined window of time at the beginning of the pupaldevelopment of the host insect (Anderson and Cervenka2002). An example of the practical application of thesewasps involved a case where the early colonizers,individuals of the blowfly Protophormia terraenovae,had finished their development and already left the scenebut adults of the parasitoid Nasonia vitripennis (Hymen-optera: Pteromalidae) were just about to emerge. Thesewasps need, at a constant temperature of 25C, 350accumulated degree days, equating to about 14 days, toreach adulthood (Whiting 1967; Grassberger and Frank2003b). By contrast the host P. terraenovaeneeds about9 days at this temperature to reach the stage appropriatefor the parasitoids oviposition (Marchenko 2001; Grass-berger and Reiter 2002a). It can therefore be assumed that
the flies had access to the body for at least about 23 daysbefore the corpse was found. The calculation of devel-opmental times for the host and the parasitoid allowed theestimation of a greater minimum postmortem intervalthan the estimated development time of Protophormiaterraenovaealone. This enabled the criminal investigatorsto disprove the testimony of a witness who claimed thathe had seen the victim alive 20 days before the corpse was
found. However, when thinking about the potentialinfluence, especially of larval parasitoids, it is importantto remember that this specialized group might also createsignificant problems for forensic entomology. Holdawayand Evans (1930) described, for example, the change indevelopmental times for Lucilia sericata after the attackof its parasitoid Alysia manducator, which resulted inpremature pupariation.
The role of freshwater and marine fauna in forensicinvestigations has received very little attention (Payneand King 1972; Nuorteva et al. 1974; Goff and Odom1987; Haskell et al 1989; Catts and Goff 1992; Vance etal. 1995; Sorg et al. 1997; Davis and Goff 2000).
Knowledge about the role of aquatic arthropods duringdecomposition is still scanty (Keiper et al. 1997;Tomberlin and Adler 1998; Hobischak and Anderson1999, 2002; Anderson 2001; Merrit and Wallace 2001;Anderson and Hobischak 2004). Compared with terres-trial habitats, decomposition in an aquatic environment iscompletely different. It occurs at a rate roughly half thatof decomposition on land, mainly due to the prevention ofinsect activity and cooler temperatures (Knight 1991).Merrit and Wallace (2001) have distinguished six de-compositional stages ranging from submerged fresh,floating decay to sunken remains. Aquatic insects offorensic importance belong to the Ephemeroptera (may-
flies), Trichoptera (caddis flies) and Diptera (true flies);the latter are mainly represented by Chironomidae(midges) and Simuliidae (black flies). However, theseinsects, unlike their terrestrial counterparts, are notobligatory saprophages, but instead use the submergedcarrion both as a food source and a breeding site. The useof these insects for estimating the time of death istherefore more difficult and depends on the season and onother conditions of the aquatic systems. No successionalinsect model exists which describes the different waves ofcolonization of a corpse in aquatic habitats (Merrit andWallace 2001).
Finally, forensic entomology may help in investiga-tions dealing with living, but ill, people by revealingneglect. The occurrence of maggots in the wounds ornatural orifices of living persons may indicate such aneglect. Estimating the age of these maggots can revealhow long the neglect has been happening (Benecke 2003).
Conclusions
Despite 150 years of use, forensic entomology is still ayoung discipline. One of the most important challengesfor the future is to combine experimental data and
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practical case work. Due to the wide variations in bioticand abiotic factors which occur at death scenes, aimprovement of the existing understanding can only beestablished through an increased number of detailed andquantified observations. Forensic entomologists are al-ways presented with the task of reconstructing the deathscene conditions as closely as possible. A model for thecalculation and handling of the data is crucial for the
credibility of this discipline.Acknowledgements Many thanks to Prof. Dr. Dietrich Mebs(Frankfurt, Germany) and Prof. Dr. Richard Wall (Bristol, UK) fortheir helpful comments on the language and content of themanuscript.
References
Allen JC (1976) A modified sine wave method for calculating daydegrees. Environ Entomol 5:388396
Amendt J, Krettek R, Niess C, Zehner R, Bratzke H (2000)Forensic entomology in Germany. Forensic Sci Int 113:309314
Ames C, Turner B (2003) Low temperature episodes in develop-ment of blowflies: implications for postmortem interval esti-mation. Med Vet Entomol 17:178186
Anderson GS (1995) The use of insects in death investigations: ananalysis of cases in British Columbia over a five year period.Can Soc Forensic J 28:277292
Anderson GS (2000) Minimum and maximum development rates ofsome forensically important Calliphoridae (Diptera). J ForensicSci 45:82432
Anderson GS (2001) Succession on carrion and its relationship todetermining time of death. In: Byrd JH, Castner JL (eds)Forensic entomology: the utility of arthropods in legal inves-tigations. CRC, Boca Raton, Fla., pp 143175
Anderson GS, Cervenka VJ (2002) Insects associated with thebody: their use and analyses. In: Haglund WD, Sorg MH (eds)Advances in forensic taphonomy: method, theory and archae-
ological perspectives. CRC, Boca Raton, Fla., pp 173200Anderson GS, Hobischak NR (2004) Decomposition of carrion inthe marine environment in British Columbia, Canada. Int JLegal Med:118
Anderson GS, VanLaerhoven SL (1996) Initial studies on insectsuccession on carrion in southwestern British Columbia. JForensic Sci 41:617625
Anonymous (1814) Instruction fr die oeffentlich angestelltenAerzte und Wundaerzte in den k.k. oesterreichischen Staaten,wie sie sich bei gerichtlichen Leichenschauen zu benehmenhaben. Schoenfeld, Prague
Arnaldos I, Romera E, Garca MD, Luna A (2001) An initial studyon the succession of sarcosaprophagous Diptera (Insecta) oncarrion in the southeastern Iberian peninsula. Int J Legal Med114:156162
Ashworth JR, Wall R (1994) Responses of the sheep blowflies
Lucilia sericataandL. cuprinato odour and the development ofsemiochemical baits. Med Vet Entomol 8:303309Baskerville GL, Emin P (1969) Rapid estimation of heat accumu-
lation from maximum and minimum temperatures. Ecology50:514517
Baumgartner DL, Greenberg B (1984) The genus Chrysomya(Diptera: Calliphoridae) in the New World. J Med Entomol21:105113
Baumgartner DL, Greenberg B (1985) Distribution and medicalecology of the blow flies (Diptera: Calliphoridae) of Peru. AnnEntomol Soc Am 78:565578
Benecke M (1998) Random amplified polymorphic DNA (RAPD)typing of necrophageous insects (Diptera, Coleoptera) in
criminal forensic studies: validation and use in practice.Forensic Sci Int 98:157168
Benecke M (2003) Neglect of the elderly: cases and considerations.Proceedings of the first meeting of the European Associationfor Forensic Entomology, pp 2930
Bergeret M (1855) Infanticide, momification naturelle du cadavre.Ann Hyg Publique Med Leg 4:442452
Beyer JC, Enos WF, Stajic M (1980) Drug identification throughanalysis of maggots. J Forensic Sci 25:411412
Blankaart S (1690) Schauplatz derer Raupen, Wrm und Maden.
LeipzigBornemissza GF (1957) An analysis of arthropod succession incarrion and the effect of its decomposition on the soil fauna.Aust J Zool 5:1-12
Bourel B, Hdouin V, Martin-Bouyer L, Becart A, Tournel G,Deveaux M, Gosset D (1999) Effects of morphine in decom-posing bodies on the development ofLucilia sericata(Diptera:Calliphoridae). J Forensic Sci 44:354358
Bourel B, Fleurisse L, Hdouin V, Cailliez JC, Creusy C, Gosset D,Goff ML (2001a) Immunohistochemical contribution to thestudy of morphine metabolism in Calliphoridae larvae andimplications in forensic entomotoxicology. J Forensic Sci46:596599
Bourel B, Tournel G, Hdouin V, Deveaux M, Goff ML, Gosset D(2001b) Morphine extraction in necrophagous insects remainsfor determining ante-mortem opiate intoxication. Forensic Sci
Int 120:127131Bourel B, Tournel G, Hedouin V, Gosset D (2004) Entomofauna ofburied bodies in Northern France. Int J Legal Med:118
Braack LEO (1981) Visitation patterns of principal species of theinsect complex at carcasses in the Kruger National Park.Koedoe 24:3349
Byrd JH, Butler JF (1998) Effects of temperature on Sarcophagahaemorrhoidalis(Diptera: Sarcophagidae) development. J MedEntomol 35:694698
Campobasso CP, Introna F (2001) The forensic entomologist in thecontext of the forensic pathologists role. Forensic Sci Int120:132139
Campobasso CP, Di Vella G, Introna F (2001) Factors affectingdecomposition and Diptera colonization. Forensic Sci Int120:1827
Campobasso CP, Gherardi M, Caligara M, Sironi L, Introna F
(2004) Drug analysis in blowfly larvae and in human tissues: acomparative study. Int J Legal Med:118Carlson DA, Geden CJ, Bernier UR (1999) Identification of pupal
exuviae ofNasonia vitripennis and Muscidifurax raptorellusparasitoids using cuticular hydrocarbons. Biol Control 15:97106
Carvalho LML, Linhares XL (2001) Seasonality of insect succes-sion and pig carcass decomposition in a natural forest area insoutheastern Brazil. J Forensic Sci 46:604608
Caterino MS, Cho S, Sperling FA (2000) The current state of insectmolecular systematics: a thriving Tower of Babel. Annu RevEntomol 45:1-54
Catts EP, Goff ML (1992) Forensic entomology in criminalinvestigations. Annu Rev Entomol 37:253272
Catts EP, Haskell NH (1990) Entomology and death: a proceduralguide. Joyces Print Shop, Clemson, USA
Clark MA, Worrell MB, Pless JE (1997) Postmortem changes insoft tissues. In: Haglund WD, Sorg MH (eds) Forensictaphonomy: the postmortem fate of human remains. CRC,Boca Raton, Fla., pp 151170
Clery JM (2001) Stability of prostate specific antigen (PSA), andsubsequent Y-STR typing, of Lucilia (Phaenicia) sericata(Meigen) (Diptera: Calliphoridae) maggots reared from asimulated postmortem sexual assault. Forensic Sci Int120:7276
Davies L (1990) Species composition and larval habitats of blowfly (Calliphoridae) populations in upland areas in England andWales. Med Vet Entomol 4:6188
61
8/10/2019 Naturwissenschaften (2004) 91.5165.pdf
12/15
Davies L, Ratcliffe GG (1994) Development rates of some pre-adult stages in blowflies with reference to low temperatures.Med Vet Entomol 8:245254
Davis JB, Goff ML. (2000) Decomposition patterns in terrestrialand intertidal habitats on Oahu Island and Coconut Island,Hawaii. J Forensic Sci 45:836842
Deonier CC (1940) Carcass temperatures and their relation towinter blowfly populations and activity in the Southwest. JEcon Entomol 33:166170
Dillon N, Austin AD, Bartowsky E (1996) Comparison of
preservation techniques for DNA extraction from hymenopter-ous insects. Insect Mol Biol 5:2124Erzinclioglu YZ (1983) The application of entomology to forensic
medicine. Med Sci Law 23:5763Erzinclioglu YZ (1990) On the interpretation of maggot evidence in
forensic cases. Med Sci Law 30:6566Erzinclioglu YZ (1996) Blowflies. Richmond Publishing, Slough,
UKFabritius K, Klunker R (1991) Die Larven- und Puparienparasitoide
von synanthropen Fliegen in Europa. Merkbl Angew Para-sitenkd Schdlingsbekmpf 32:124
Faucherre J, Cherix D, Wyss C (1999) Behavior of Calliphoravicina (Diptera, Calliphoridae) under extreme conditions. JInsect Behav 12:687690
Fisher P, Wall R, Ashworth JR (1998) Attraction of the sheepblowfly, Lucilia sericata (Diptera: Calliphoridae) to carrion
bait in the field. Bull Entomol Res 88:611616Fuller ME (1934) The insect inhabitants of carrion: a study inanimal ecology. Council for Scientific and Industrial Research,Bulletin 82
Gagliano-Candela R, Aventaggiato L (2001) The detection of toxicsubstances in entomological specimens. Int J Legal Med114:197203
Gasser RB, Chilton NB (2001) Applications of single-strandconformation polymorphism (SSCP) to taxonomy, diagnosis,population genetics and molecular evolution of parasiticnematodes. Vet Parasitol 101:201213
Geden CJ (1997) Development models for the filth fly parasitoidsSpalangia gemina, S. cameroni, and Muscidifurax raptor(Hymenoptera: Pteromalidae) under constant and variabletemperatures. Biol Control 9:185192
Geden CJ, Bernier UR, Carlson DA, Sutton BD (1998) Identifi-
cation of Muscidifurax spp., parasitoids of muscoid flies, bycomposition patterns of cuticular hydrocarbons. Biol Control12:200207
Goddard J, Lago PK (1985) Notes on blowfly (Diptera: Callipho-ridae) succession on carrion in Northern Mississippi. J EntomolSci 20:312317
Godfray HCJ (1994) Parasitoids: behavioral and evolutionaryecology. Princeton University Press, Princeton, N.J.
Goff ML (1991) Comparison of insect species associated withdecomposing remains recovered inside dwellings and outdoorson the island of Oahu, Hawaii. J Forensic Sci 3:748753
Goff ML, Flynn MM (1991) Determination of postmortem intervalby arthropod succession: a case study from the HawaiianIsland. J Forensic Sci 36:607614
Goff ML, Lord WD (1994) Entomotoxicology: a new area forforensic investigation. Am J Forensic Med Pathol 15:5157
Goff ML, Lord WD (2001) Entomotoxicology: insects as toxico-logical indicators and the impact of drugs and toxins on insectdevelopment. In: Byrd JH, Castner JL (eds) Forensic entomol-ogy: the utility of arthropods in legal investigations. CRC, BocaRaton, Fla., pp 331340
Goff ML, Odom CB (1987) Forensic entomology in the HawaiianIslands: three case studies. Am J Forensic Med Pathol 8:4550
Goff ML, Odom CB, Early M (1986) Estimation of postmorteminterval by entomological techniques: a case study from Oahu,Hawaii. Bull Soc Vector Ecol 11:242246
Goff ML, Omori AI, Goodbrod JR (1989) Effect of cocaine intissues on the rate of development of Boettcherisca peregrina(Diptera: Sarcophagidae). J Med Entomol 26:9193
Goff ML, Brown WA, Hewadikaram KA, Omori AI (1991) Effectsof heroin in decomposing tissues on the development rate ofBoettcherisca peregrina(Diptera: Sarcophagidae) and implica-tions of this effect on estimation of postmortem intervals usingarthropod development patterns. J Forensic Sci 36:537542
Goff ML, Miller ML, Paulson JD, Lord WD, Richards E, Omori AI(1997) Effects of 3,4-methylendioxymethamphetamine in de-composing tissues on the development of Parasarcophagaruficornis(Diptera: Sarcophagidae) and detection of the drug inpostmortem blood, liver tissue, larvae, and puparia. J Forensic
Sci 42:276280Grassberger M, Frank C (2003a) Initial study of arthropodsuccession on pig carrion in a central European urban habitat.J Med Entomol:40
Grassberger M, Frank C (2003b) Temperature-dependent develop-ment of the parasitic wasp Nasonia vitripennisand its forensicimplications. Med Vet Entomol 17:257262
Grassberger M, Reiter C (2001) Effect of temperature on Luciliasericata (Diptera: Calliphoridae) development with specialreference to the isomegalen- and isomorphen-diagram. ForensicSci Int 120:3236
Grassberger M, Reiter C (2002a) Effect of temperature ondevelopment of the forensically important holarctic blowflyProtophormia terraenovae(Robineau-Desvoidy) (Diptera: Cal-liphoridae). Forensic Sci Int 128:177182
Grassberger M, Reiter C (2002b) Effect of temperature on
development of Liopygia (=Sarcophaga) argyrostoma (Robi-neau-Desvoidy) (Diptera: Sarcophagidae) and its forensicimplications. J Forensic Sci 47:13321336
Greenberg B (1984) Two cases of human myiasis caused byPhaenicia sericata (Diptera: Calliphoridae) in Chicago areahospitals. J Med Entomol 21:615
Greenberg B (1985) Forensic entomology: case studies. BullEntomol Soc Am 31:2528
Greenberg B (1990) Nocturnal oviposition behaviour of blow flies(Diptera: Calliphoridae). J Med Entomol 27:807810
Greenberg B (1991) Flies as forensic indicators. J Med Entomol28:565577
Greenberg B, Kunich JC (2002) Entomology and the law: flies asforensic indicators. Cambridge University Press, Cambridge
Gunatilake K, Goff ML (1989) Detection of organophosphatepoisoning in a putrefying body by analyzing arthropod larvae. J
Forensic Sci 34:714716Hall MJR (1995) Trapping the flies that cause myiasis: theirresponses to host-stimuli. Ann Trop Med Parasitol 89:333357
Hall RD (2001) Perceptions and status of forensic entomology In:Byrd JH, Castner JL (eds) Forensic entomology: the utility ofarthropods in legal investigations. CRC, Boca Raton, Fla., pp115
Hall MJR, Farkas R, Kelemen F, Hosier MJ, El-Khoga JM (1995)Orientation of agents of wound myiasis to hosts and artificialstimuli in Hungary. Med Vet Entomol 9:7784
Harvey M, Dadour I, Gaudieri S (2003) Mitochondrial DNAcytochrome oxidase I gene: potential for distinction betweenimmature stages of some forensically important fly species(Diptera) in western Australia. Forensic Sci Int 131:134139
Haskell NH, McShaffrey DG, Hawley DA, Williams RE, Pless JE(1989) Use of aquatic insects in determining submersioninterval. J Forensic Sci 34:622632
Haskell NH, Hall RD, Cervenka VJ, Clark MA (1997) On the body:insects life stage presence, their postmortem artifacts. In:Haglund WD, Sorg MH (eds) Forensic taphonomy: thepostmortem fate of human remains. CRC, Boca Raton, Fla.,pp 415448
Hauser G (1926) Ein Beitrag zum Madenfra an menschlichenLeichen. Dtsch Z Gesamte Gerichtl Med 7:179192
Hedouin V, Bourel B, Becart A, Tournel G, Deveaux M, Goff ML,Gosset D (2001) Determination of drug levels in larvae ofProtophormia terraenovae and Calliphora vicina (Diptera:Calliphoridae) reared on rabbit carcasses containing morphine.J Forensic Sci 46:1214
62
8/10/2019 Naturwissenschaften (2004) 91.5165.pdf
13/15
Henge C, Madea B, Knight B, Nokes L, Krompecher T (1995)The estimation of the time since death in the early postmorteminterval. Arnold, London
Henge C, Althaus L, Bolt J, Freislederer A, Haffner HT, HengeCA, Hoppe B, Schneider V (2000a) Experiences with acompound method for estimating the time since death. I.Rectal temperature nomogram for time since death. Int J LegalMed 113:303319
Henge C, Althaus L, Bolt J, Freislederer A, Haffner HT, HengeCA, Hoppe B, Schneider V (2000b) Experiences with a
compound method for estimating the time since death. II.Integration of non-temperature-based methods. Int J Legal Med113:320331
Higley LG, Haskell NH (2001) Insect development and forensicentomology. In: Byrd JH, Castner JL (eds) forensic entomol-ogy: the utility of arthropods in legal investigations. CRC, BocaRaton, Fla., pp 287302
Hobischak NR, Anderson GS (1999) Freshwater-related deathinvestigations in British Columbia in 19951996, a review ofcoroners cases. Can Soc Forensic Sci 32:97106
Hobischak NR, Anderson GS (2002) Time of submergence usingaquatic invertebrate succession and decompositional changes. JForensic Sci 47:142151
Holdaway FG, Evans AC (1930) Parasitism a stimulus to pupation:Alysia manducator in relation to the host Lucilia sericata.Nature 125:598599
Horoszkiewicz S von (1902) Casuistischer Beitrag zur Lehre vonder Benagung der Leichen durch Insekten. VierteljahresschrGerichtl Med 23:235239
Introna F, Campobasso CP (2000) Forensic dipterology. In: Papp L,Darvas B (eds) Contributions to a manual of palaearctic diptera.1. General and applied dipterology. Science Herald, Budapest,pp 793846
Introna F, Altamura BM, DellErba A, Dattoli V (1989) Time sincedeath definition by experimental reproduction of Luciliasericatacycles in growth cabinet. J Forensic Sci 34:478480
Introna F, Lo Dico C, Caplan YH, Smialek JE (1990) Opiateanalysis in cadaveric blowfly larvae as an indicator of narcoticintoxication. J Forensic Sci 35:118122
Introna F, Suman TW, Smialek JE (1991) Sarcosaprophagous flyactivity in Maryland. J Forensic Sci 36:238243
Introna F, Campobasso CP, Di Fazio A (1998) Three case studies in
forensic entomology from southern Italy. J Forensic Sci43:210214Introna F, Campobasso CP, Goff ML (2001) Entomotoxicology.
Forensic Sci Int 120:4247Junqueira ACM, Lessinger AC, Azeredo-Espin AML (2002)
Methods for the recovery of mitochondrial DNA sequencesfrom museum specimens of myiasis-causing flies. Med VetEntomol 16:3945
Kamal AS (1958) Comparative study of thirteen species ofsarcosaprophagous Calliphoridae and Sarcophagidae (Diptera).I. Bionomics. Ann Entomol Soc Am 51:261270
Kaneshrajah G, Turner B (2004) Calliphora vicina larvae grow atdifferent rates on different body tissues. Int J Legal Med:118
Keiper JB, Chapman EG, Foote BA (1997) Midge larvae (Diptera:Chironomidae) as indicators of postmortem submersion intervalof carcasses in a woodland stream: a preliminary report. JForensic Sci. 42:10741079
Kintz P, Tracqui A, Mangin P (1990) Toxicology and fly larvae ona putrefied cadaver. J Forensic Sci Soc 30:243246
Kintz P, Tracqui A, Mangin P (1994) Analysis of opiates in flylarvae sampled on a putrefied cadaver. J Forensic Sci Soc34:9597
Knight B (1991) Forensic pathology. Edward Arnold, LondonKrahmer FL (1857) Handbuch der gerichtlichen Medizin. 2. Aufl.
BerlinLaSalle J, Gauld ID (1991) Parasitic Hymenoptera and the
biodiversity crisis. Redia 74:315334Leclercq M (1983) Entomologie et mdecine lgale; datation de la
mort, observation indite. Rev Med Liege 38:735738
Leclercq J, Leclercq M (1948) Donnes bionomiques pourCalliphora erythrocephala (Meigen) et cas dapplication lamedecine lgale. Bull Soc Entomol Fr 53:101103
Lord WD, Catts EP, Scarboro DA, Hadfield DB (1986) The greenblow fly, Lucilia illustris (Meigen), as an indicator of humanpostmortem interval: a case of homicide from Fort Lewis,Washington. Bull Soc Vector Ecol 11:271275
Loxdale HD, Lushai G (1998) Molecular markers in entomology.Bull Entomol Res 88:577600
Malgorn Y, Coquoz R (1999) DNA typing for identification of
some species of Calliphoridae: an interest in forensic entomol-ogy. Forensic Sci Int 102:111119Mann RW, Bass WM, Meadows L (1990) Time since death and
decomposition of the human body: variables and observationsin case and experimental field studies. J Forensic Sci 35:103111
Marchenko MJ (1980) Classifying of cadaveric entomofauna.Biology of flies: the forensic medical role. Sud-Med Ekspert23:1720
Marchenko MI (1988) Medico-legal relevance of cadaver entomo-fauna for the determination of the time since death. ActaMedicinae Et Socialis Organe Officiel De LAcademie Inter-nationale De Medicine Legale Et De Medicine Sociale 38:257302.
Marchenko MJ (2001) Medicolegal relevance of cadaver entomo-fauna for the determination of time since death. Forensic Sci Int
120:89109McKnight BE (1981) The washing away of wrongs: forensicmedicine in thirteenth-century China. University of Michigan,Ann Arbor
McWatters HG, Saunders DS (1998) Maternal temperature hasdifferent effects on the photoperiodic response and duration oflarval diapause in blow fly (Calliphora vicina) strains collectedat two latitudes. Physiol Entomol 23:369375
Mgnin JP (1894) La faune des cadavres: application de lento-mologie a la mdecine lgale. Masson et Gauthiers-Villars,Paris
Mende LJK (1829) Ausfhrliches Handbuch der gerichtlichenMedizin fr Gesetzgeber, Rechtsgelehrte, Aerzte und Wund-aerzte, Teil 5
Merrit RW, Wallace JR (2001) The role of aquatic insects inforensic investigations. In: Byrd JH, Castner JL (eds) Forensic
entomology: the utility of arthropods in legal investigations.CRC, Boca Raton, Fla., pp 177222Miller ML, Lord WD, Goff ML, Donnelly D, McDonough ET,
Alexis JC (1994) Isolation of amitriptyline and nortriptylinefrom fly pupariae (Phoridae) and beetle exuviae (Dermestidae)associated with mummified human remains. J Forensic Sci39:13051313
Nishida K, Shinonaga S, Kano R (1986) Growth tables of fly larvaefor the estimation of postmortem intervals. Ochanomizu Med J34:924
Nolte KB, Pinder RD, Lord WD (1992) Insect larvae used to detectcocaine poisoning in a decomposed body. J Forensic Sci37:11791185
Nuorteva P (1959a) Studies on the significance of flies in thetransmission of poliomyelitis. III. The composition of the blowfly fauna and the activity of the flies in relation to the weatherduring the epidemic season of poliomyelitis in south Finland.Ann Entomol Fenn 25:137162
Nuorteva P (1959b) Studies on the significance of flies in thetransmission of poliomyelitis. IV. The composition of the blowfly fauna in different part of Finland during 1958. Ann EntomolFenn 25:137162
Nuorteva P (1965) The flying activity of blowflies (Diptera,Calliphoridae) in subarctic conditions. Ann Entomol Fenn31:242245
Nuorteva P (1977) Sarcosaprophagous insects as forensic indica-tors. In: Tedeshi GC, Eckert WG, Tedeshi LG (eds) Forensicmedicine: a study in trauma and environmental hazards, vol 2,Saunders, Philadelphia, pp 10721095
63
8/10/2019 Naturwissenschaften (2004) 91.5165.pdf
14/15
Nuorteva P, Nuorteva SL (1982) The fate of mercury in sarcos-aprophagous flies and in insects eating them. Ambio 11:3437
Nuorteva P, Isokoski M, Laiho K (1967) Studies on the possibilitiesof using blowflies (Dipt.) as medicolegal indicators in Finland.Ann Entomol Fenn 33:217225
Nuorteva P, Schumann H, Isokoski M, Laiho K (1974) Studies onthe possibilities of using blowflies (Diptera: Calliphoridae) asmedicolegal indicators in Finland. Ann Entomol Fenn 40:7074
OBrien C, Turner B (2004) Impact of paracetamol on thedevelopment of Calliphora vicina larval development. Int J
Legal Med:118Orfila MJB, Lesueur CA (1831) Trait des exhumations juridiques.Paris
Pbo S (1989) Ancient DNA: extraction, characterization, molec-ular cloning, and enzymatic amplification. Proc Natl Acad SciUSA 86:19391943
Pbo S, Higushi RG, Wilson AC (1989) Ancient DNA and thepolymerase chain reaction. J Biol Chem 264:97099712
Payne JA (1965) A summer carrion study of the baby pig Sus scrofaLinnaeus. Ecology 46:592602
Payne JA, King EW (1972) Insect succession and decomposition ofpig carcases in water. J Ga Entomol Soc 7:153162
Payne JA, King EW, Beinhart G (1968) Arthropod succession anddecomposition of buried pigs. Nature 219:11801181
Pien K, Marichal M, Grootaert P, De Boeck G, Samyn N, BoonenT, Vits K, Wood M, Morris M (2004) The detection of
nordiazepam and its metabolite oxazepam in one singlepostfeeding larva and puparium of Calliphora vicina (Diptera:Calliphoridae) using the LC/MS-MS. Int J Legal Med:118
Povoln D, Verves Y (1997) The flesh-flies of central Europe.Spixiana Suppl 24:1260
Price PW (1997) Insect ecology. Wiley, New YorkRath PM, Ansorg R (2000) Identification of medically important
Aspergillus species by single strand conformational polymor-phism (SSCP) of the PCR-amplified intergenic spacer region.Mycoses 43:381386
Reed HB (1958) A study of dog carcass communities in Tennessee,with special references to the insects. Am Midl Nat 59:213245
Reinhard H (1882) Beitrge zur Grberfauna. Verh Kaiserl-KniglZool-Bot Ges Wien 31:207210
Reiter C (1984) Zum Wachstumsverhalten der Maden der blauenSchmeifliege Calliphora vicina. Z Rechtsmed 91:295308
Reiter C, Wolleneck G (1982) Bemerkungen zur Morphologieforensisch bedeutsamer Fliegenmaden. Z Rechtsmed 89:197206
Reiter C, Wolleneck G (1983) Zur Artbestimmung der Madenforensisch bedeutsamer Schmeifliegen. Z Rechtsmed 90:309316
Richards EN, Goff ML (1997) Arthropod succession on exposedcarrion in three contrasting tropical habitats on Hawaii Island,Hawaii. J Med Entomol 34:328339
Rodriguez WC, Bass WM (1983) Insect activity and its relationshipto decay rates of human cadavers in East Tennessee. J ForensicSci 28:423432
Rodriguez WC, Bass WM (1985) Decomposition of buried bodiesand methods that may aid in their location. J Forensic Sci30:836852
Sadler DW, Fuke C, Court F, Pounder DJ (1995) Drug accumu-lation and elimination in Calliphora vicinalarvae. Forensic SciInt 71:191197
Sadler DW, Robertson L, Brown G, Fuke C, Pounder DJ (1997a)Barbiturate and analgesics in Calliphora vicina larvae. JForensic Sci 42:481485
Sadler DW, Chuter G, Senevematne C, Pounder DJ (1997b)Commentary on Sadler DW, Robertson L, Brown G, Fuke C,Pounder DJ, Barbiturates and analgesics in Calliphora vicinalarvae. J Forensic Sci 42:12141215
Sadler DW, Richardson J, Haigh S, Bruce G, Pounder DJ (1997c)Amitryptiline accumulation and elimination in Calliphoravicinalarvae. Am J Forensic Med Pathol 18:397403
Sambrook J, Russel DW (2001) Molecular cloning: a laboratorymanual. Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y.
Saunders DS (1975) Manipulation of the length of the sensitiveperiod, and the induction of pupal diapause in the flesh-flySarcophaga argyrostoma. J Entomol 50:107118
Schmitz H (1928) Phoriden in doodkisten. Natuurhist Maandbl17:150153
Schneider P (1936) Leichenzerstrung durch Madenfra. Wie langelag die Leiche im Gebsch? Arch Kriminol 98:216221
Schoenly K, Reid W (1987) Dynamics of heterotrophic successionin carrion arthropod assemblages: discrete series or a contin-uum of change? Oecologia 73:192202
Schoenly K, Goff ML, Wells JD, Lord WD (1996) Quantifyingstatistical uncertainty in succession-based entomological esti-mates of the postmortem interval in death scene investigations:a simulation study. Am Entomol 42:106112
Schumann H (1971) Die Gattung Lucilia (Goldfliegen). MerkblAngew Parasitenkd Schdlingsbekmpf 18:120
Schrder H, Klotzbach H, Elias S, Augustin C, Pueschel K (2003)Use of PCR-RFLP for differentiation of calliphorid larvae(Diptera, Calliphoridae) on human corpses. Forensic Sci Int132:7681
Shean BSL, Messinger L, Papworth (1993) Observations ofdifferential decomposition on sun exposed vs. shaded pigcarrion in coastal Washington State. J Forensic Sci 38:938949
Sherman RA (2000) Wound myiasis in urban and suburban UnitedStates. Arch Intern Med 160:20042014Shimko N, Liu L, Lang BF, Burger, G (2001) GOBASE: the
organelle genome database. Nucleic Acids Res 29:128132Simon C, Frati F, Beckenbach A, Crespi B, Liu H, Flook P (1994)
Evolution, weighting and phylogenetic utility of mitochondrialgene sequences and a compilation of conserved polymerasechain reaction primers. Ann Entomol Soc Am 87:651701
Singh D, Bharti M (2001) Further observations on the nocturnaloviposition behaviour of blowflies (Diptera: Calliphoridae).Forensic Sci Int 120:124126
Smith KE, Wall R (1997a) Asymmetric competition between larvaeof the blowflies Calliphora vicina and Lucilia sericata incarrion. Ecol Entomol 22:467474
Smith KE, Wall R (1997b) The use of carrion as breeding sites bythe blowfly Lucilia sericata and other Calliphoridae. Med Vet
Entomol 11:3844.Smith KGV (1986) A manual of forensic entomology. BritishMuseum, London
Sorg MH, Dearborn JH, Monahan EI, Ryan HF, Sweeney KG,David E (1997) Forensic taphonomy in marine contexts. In:Haglund WD, Sorg MH (eds) Forensic taphonomy: thepostmortem fate of human remains. CRC, Boca Raton, Fla.,pp 567604
Sperling FA, Anderson GS, Hickey DA (1994) A DNA-basedapproach to the identification of insect species used forpostmortem interval estimation. J Forensic Sci 39:418427
Stafford F (1971) Insects of a medieval burial. Sci Anthropol 7:610
Stevens JR, Wall R (1996) Species, sub-species and hybridpopulations of the blowflies Lucilia cuprina and Luciliasericata (Diptera: Calliphoridae). Proc R Soc Lond B263:13351341
Stevens JR, Wall R (2001) Genetic relationships between blowflies(Calliphoridae) of forensic importance. Forensic Sci Int120:11623
Stevens JR, Wall R, Wells JD (2002) Paraphyly in Hawaiian hybridblowfly populations and the evolutionary history of anthropo-philic species. Insect Mol Biol 11:141148
Tessmer JW, Meek CL, Wright VL (1995) Circadian patterns ofoviposition by necrophilous flies (Diptera: Calliphoridae).Southwest Entomol 24:439445
Tomberlin JK, Adler PH (1998) Seasonal colonization anddecomposition of rat carrion in water and on land in an openfield in South Carolina. J Med Entomol 35:704709
64
8/10/2019 Naturwissenschaften (2004) 91.5165.pdf
15/15
Tracqui A, Keyser-Tracqui C, Kintz P, Ludes B (2004) Entomo-toxicology for the forensic toxicologist: much ado aboutnothing? Int J Legal Med:118
Vance GM, VanDyk JK, Rowley WA (1995) A device for samplingaquatic insects associated with carrion in water. J Forensic Sci40:479482
VanLaerhoven SL, Anderson GS (1999) Insect succession onburied carrion in two biogeoclimatic zones of British Columbia.J Forensic Sci 44:3144
Vincent S, Vian JM, Carlotti MP (2000) Partial sequencing of the
cytochrome oxydase b subunit gene I: a tool for the identifi-cation of European species of blow flies for postmorteminterval estimation. J Forensic Sci 45:820823
Vinogradova EB (1986) Geographic variation and ecologicalcontrol of diapause in flies. In: Taylor F, Karban R (eds) Theevolution of insect life-cycles. Springer, Berlin Heidelberg NewYork, pp 3547
Vinogradova EB (1991) Diapause in flies and its control (inRussian with English summary). Proc ZIN RAS 214, St.Petersburg
Vinogradova EB, Marchenko MJ (1984) The use of temperatureparameters of fly growth in the medicolegal practice. Sud-MedEkspert 27:1619
Vinogradova EB, Zinovjeva KB (1972) Maternal induction oflarval diapause in the blowfly Calliphora vicina. J InsectPhysiol 18:24012409
Wall R, Fisher P (2001) Visual and olfactory cue interaction inresource-location by the blowfly, Lucilia sericata. PhysiolEntomol 26:212218
Wall R, Warnes ML (1994) Responses of the sheep blowfly Luciliasericata to carrion odour and carbon dioxide. Entomol ExpAppl 73:239246
Wallman JF, Donnellan SC (2001) The utility of mitochondrialDNA sequences for the identification of forensically importantblowflies (Diptera: Calliphoridae) in southeastern Australia.Forensic Sci Int 120:6067
Watson EJ, Carlton CE (2003) Spring succession of necrophilousinsects on wildlife carcasses in Louisiana. J Med Entomol40:338347
Weismann A (1864) Die nachembryonale Entwicklung der Mus-ciden nach Beobachtungen an Musca vomitoria und Sarco-phaga carnaria. Z Wiss Zool 14:187336
Wells JD, King L (2001) Incidence of precocious egg developmentin flies of forensic importance (Calliphoridae). Pan-Pac Ento-mol 77:235239
Wells JD, LaMotte LR (1995) Estimating maggot age from weightusing inverse prediction. J Forensic Sci 40:585590
Wells JD, Sperling FA (1999) Molecular phylogeny ofChrysomyaalbiceps and C. rufifacies (Diptera: Calliphoridae). J MedEntomol 36:222226
Wells JD, Sperling FA (2000) A DNA-based approach to theidentification of insect species used for postmortem intervalestimation and partial sequencing of the cytochrome oxydase bsubunit gene I: a tool for the identification of European speciesof blow flies for postmortem interval estimation. J Forensic Sci
45:13581359Wells JD, Sperling FA (2001) DNA-based identification offorensically important Chrysomyinae (Diptera: Calliphoridae).Forensic Sci Int 120:110115
Wells JD, Byrd JH, Tantawi TI (1999) Key to third-instarchrysomyinae (Diptera: Calliphoridae) from carrion in thecontinental United States. J Med Entomol 36:638641
Wells JD, Pape T, Sperling FAH (2001a) DNA based identificationand molecular systematics of forensically important Sarco-phagidae (Diptera). J Forensic Sci 46:8791
Wells JD, Introna F Jr, Di Vella G, Campobasso CP, Hayes J,Sperling FA (2001b) Human and insect mitochondrial DNAanalysis from maggots. J Forensic Sci 46:685687
Whiting AR (1967) The biology of the parasitic wasp Mormoniellavitripennis (Walker). Q Rev Biol 42:333406
Wigglesworth VB (1972) The principles of insect physiology.
Chapman and Hall, LondonWilliams H (1984) A model for the aging of fly larvae in forensicentomology. Forensic Sci Int 25:191199
Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV(1990) DNA polymorphisms amplified by arbitrary primers areuseful as genetic markers. Nucleic Acids Res 18:65316535
Yovanovich P (1888) Entomologie applique la Mdecine lgale.Olliver-Henry, Paris
Zehner R, Amendt J, Krettek R (2004a) STR typing of human DNAfrom fly larvae fed on decomposing bodies. J Forensic Sci (inpress)
Zehner R, Amendt J, Schtt S, Sauer S, Krettek R, Povoln D(2004b) Genetic identification of forensically important fleshflies (Diptera: Sarcophagidae). Int J Legal Med:118
Zumpt F (1965) Myiasis in man and animals in the old world.Butterworths, London
65